摘要
目的研究洋地黄毒苷(Digitoxin)在体外对人CD4+T细胞产生IL-17A和IFN-γ的影响及其机制。方法人PBMC用抗CD3和抗CD28抗体共刺激或在诱导Th17分化的条件下,同时加或不加Digitoxin,用酶联免疫吸附法检测培养上清中IL-17A和IFN-γ的产生。利用流式细胞术检测细胞因子IL-17A、IFN-γ以及转录因子RORγt的表达。利用Western blotting检测Digitoxin对RORγt表达的影响。结果 Digitoxin对人IL-17A有显著的剂量依赖性抑制作用,并且在刺激的早期(第0小时和第6小时)加入Digitoxin能显著地抑制IL-17A的产生,但不影响IFN-γ的产生,而晚期(第12小时和第24小时)加入Digitoxin对IL-17A的产生没有明显地抑制作用。进一步研究发现,Digitoxin抑制Th17细胞的分化,调控IL-17A的转录因子RORγt的表达。结论本实验表明,Digitoxin通过早期调控RORγt的表达来抑制IL-17A的产生,但不抑制IFN-γ的产生,提示Digitoxin除了对心血管作用外,也可以抑制炎症性疾病和自身免疫性疾病的发生发展。
To study the effect of Digitoxin on the production of IL-17A and IFN-γ by human CD4＋ T cells in vitro and investigate its mechanism, PBMCs were stimulated with anti-CD3 plus anti-CD28 and naive CD3＋CD4＋ T cells were differentiated into Th17 cells in the presence or absence of Digitoxin. Then the levels of IL-17A and IFN-γ in culture supernatants were detected by enzyme-linked immunosorbent assay （ELISA）; the expression of cytokines （IL-17A and IFN-γ） and transcription factor RORγt were analyzed by fluorescence activated cell sorter （FACS） or Western blotting. We found that Digitoxin significantly suppressed the production of IL-17A in a dose- dependent manner, and the suppressive effect was more remarkable when Digitoxin was added at early timepoints （0 h and 6 h）, whereas IL-17A was not inhibited at later timepoints （12 h and 24 h）. However, Digitoxin had no significantly effect on the production of IFN-γ. Further studies revealed that Digitoxin suppressed the expression of RORγt, which plays a predominating role in regulating IL-17A production. In conclusion, Digitoxin could suppress the differentiation of Th17 cells and the production of IL-17A via regulating the expression of RORγt, but has no effect on the production of IFN-γ. Excepting the effects on cardiovascular diseases, Digitoxin might be used to treat inflammatory and autoimmunity diseases.
出处
《免疫学杂志》
CAS
CSCD
北大核心
2014年第6期519-524,共6页
Immunological Journal
基金
广东省科技计划项目(2012B040304008)
国家自然科学基金资助项目(31170833)
